TRIGLYCERIDES REAGENTS

TRIGLYCERIDES

Clinical Chemistry Reagents

Enzymatic colorimetric method
ENDPOINT

PRINCIPLE

The method is based on the enzymatic hydrolysis of serum or plasma triglyceride to glycerol and free fatty acids (FFA) by lipoprotein lipase (LPL). The glycerol is phosphorylated by adenosine triphosphate (ATP) in the presence of glycerolkinase (GK) to form glycerol-3-phosphate (G-3-P) and adenosine diphosphate (ADP).

G-3-P is oxidized by glycerophosphate oxidase (GPO) to form dihydroxyacetone phosphate (DHAP) and hydrogen peroxide.

A red chromogen is produced by the peroxidase (POD) catalyzed coupling of 4-aminoantipyrine (4-AA) and phenol with hydrogen peroxide (H2O2), proportional to the concentration of triglyceride in the sample.

REAGENT COMPOSITION

4 x 100 ml

R1 Monoreagent. PIPES buffer 50 mmol/L pH 6.8, LP greater than or equal to 12
KU/L, GK greater than or equal to 1 KU/L, GPO greater than or equal to 10 KU/L, ATP 2.0 mmol/L,
Mg2+ 40 mmol/L, POD greater than or equal to 2.5 KU/L, 4-AA 0.5 mmol/L,
phenol 3 mmol/L, non-ionic tensioactives 2 g/L (w/v).
Biocides.
CAL Triglycerides standard. Glycerol 2.26 mmol/L, equivalent
to 200 mg/dL of glycerol trioleate. Secondary standard.
Concentration value is traceable to Standard Reference
Material 909b.

STORAGE AND STABILITY
Store at 2-8ºC

REAGENT PREPARATION
The Monoreagent and the Standard are ready-to-use.

SAMPLES
Serum, EDTA or heparinized plasma free of hemolysis. Remove
from cells within 2 hours of venipuncture. Analyze samples
immediately or refrigerate. Stable for 1 week at 4-8ºC.

MATERIALS REQUIRED

• Photometer or colorimeter capable of measuring absorbance at 500 ± 20 nm.
• Constant temperature incubator set at 37ºC.
• Pipettes to measure reagent and samples.

TRIGLYCERIDES Reagents per box